长裂苦苣菜水提取物对肺癌细胞A549增殖和凋亡的影响
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摘要
为了探讨长裂苦苣菜水提取物对肺癌细胞A549的影响,本研究用不同浓度的长裂苦苣菜水提取物作用于体外培养的A549细胞,从细胞形态(显微镜观察)、细胞增殖活力(CCK-8法)、细胞凋亡(Annexin V-FITC/PI染色)、线粒体膜电位(JC-1染色)和细胞内活性氧水平(荧光探针DCFH-DA染色)几个方面检测长裂苦苣菜水提取物对体外培养的A549细胞的影响。当处理浓度达到1 mg/mL作用细胞48 h后,细胞出现明显皱缩的凋亡形态学特征,与对照组相比处理组细胞增殖活力明显下降(P<0.05)。进一步用流式细胞仪检测发现≥1mg/mL的处理浓度作用细胞48 h后细胞凋亡率显著提高(P<0.05),同时检测到处理组细胞的膜电位都明显下降,细胞内活性氧水平明显上升,都呈剂量依赖关系。这些结果表明长裂苦苣菜水提取物可以诱导A549细胞凋亡,并抑制其生长和增殖,是一种潜在的预防和抑制肿瘤生长的药食两用植物。
To illustrate the effects of Sonchus branchyotus DC( SBDC) aqueous extracts on human lung cancer A549 cells,assays,including morphologic changes observed by microscope,cell proliferation vitality tested by cell growth assay using cck8 kit,the apoptosis detected by Flow Cytometry,mitochondrial membrane potential assessed with the fluorescent probe JC-1 and reactive oxygen species formation measured by DCFH-DA staining,were performed. Observed with microscope,cells treated with high dose(≥1 mg /mL) of SBDC were shrunken and the morphological characteristics of apoptosis was obvious compared with the control group. Also,cell proliferation was significantly inhibited compared with the control group(P < 0. 05). Furthermore,flow cytometry analysis revealed the apoptosis cells treated with high dose of SBDC(≥1mg /mL) for 48 hours were significantly increased compared with the control group. Finally,it was found in all treatment groups that the significant decrease of cell mitochondrial membrane potential and the increase of reactive oxygen species level. These results demonstrated that SBDC may induce human lung cancer A549 cells apoptosis and inhibit the cells growth and proliferation. SBDC is a potential edible herb for cancer prevention and treatment.
To illustrate the effects of Sonchus branchyotus DC( SBDC) aqueous extracts on human lung cancer A549 cells,assays,including morphologic changes observed by microscope,cell proliferation vitality tested by cell growth assay using cck8 kit,the apoptosis detected by Flow Cytometry,mitochondrial membrane potential assessed with the fluorescent probe JC-1 and reactive oxygen species formation measured by DCFH-DA staining,were performed. Observed with microscope,cells treated with high dose(≥1 mg /mL) of SBDC were shrunken and the morphological characteristics of apoptosis was obvious compared with the control group. Also,cell proliferation was significantly inhibited compared with the control group(P < 0. 05). Furthermore,flow cytometry analysis revealed the apoptosis cells treated with high dose of SBDC(≥1mg /mL) for 48 hours were significantly increased compared with the control group. Finally,it was found in all treatment groups that the significant decrease of cell mitochondrial membrane potential and the increase of reactive oxygen species level. These results demonstrated that SBDC may induce human lung cancer A549 cells apoptosis and inhibit the cells growth and proliferation. SBDC is a potential edible herb for cancer prevention and treatment.
引文
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4 Green DR,Reed JC.Mitochondria and apoptosis.Science,1998,281:1309-1312.
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7 Wang J(王晶),Yi J(易静).Cancer cell killing via ROS:to increase or decrease,that is the question.Cancer Biol Ther,2008,7:1875-1884.
8 Friesen C,Fulda S,Debatin KM.Induction of CD95 ligand and apoptosis by doxorubicin is modulated by the redox state in chemosensitive and drug resistant tumor cells.Cell Death Differ,1999,6:471-480.
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2 Vilela FC,Bitencourt AD,Cabral LD,et al.Anti-inflammatory and antipyretic effects of Sonchus oleraceus in rats.J Ethnopharmacol,2010,3:737-741.
3 Thomson B,Shaw I.A comparison of risk and protective factors for colorectal cancer in the diet of New Zealand maori and non-maori.Asian Pac J Cancer Prev,2002,3:319-324.
4 Green DR,Reed JC.Mitochondria and apoptosis.Science,1998,281:1309-1312.
5 Ly JD,Grubb DR,Lawen A.The mitochondrial membrane potential[deltapsi(m)]in apoptosis;an update.Apoptosis,2003,8:115-128.
6 Trachootham D,Alexandre J,Huang P.Targeting cancer cells by ROS-mediated mechanisms:a radical therapeutic approach?Nat Rev Drug Discov,2009,8:579-591.
7 Wang J(王晶),Yi J(易静).Cancer cell killing via ROS:to increase or decrease,that is the question.Cancer Biol Ther,2008,7:1875-1884.
8 Friesen C,Fulda S,Debatin KM.Induction of CD95 ligand and apoptosis by doxorubicin is modulated by the redox state in chemosensitive and drug resistant tumor cells.Cell Death Differ,1999,6:471-480.
9 Dai J,Weinberg RS,Waxman S,et al.Malignant cells can be sensitized to undergo growth inhibition and apoptosis by arsenic trioxide through modulation of the glutathione redox system.Blood,1999,93:268-277.
10 Sun SY,Li W,Yue P,et al.Mediation of N-(4-hydoxyphenyl)retinamide induced apoptosis in human cancer cells by different mechanisms.Cancer Res,1999,59:2493-2498.